Background technology
There are some researches show that the dissolution rate of traditional powder is directly proportional with the specific grain surface of medicine is long-pending, and specific surface area and particle diameter are inversely proportional to.Therefore, the particle diameter of medicine is thin more, and then its specific surface area is big more, helps the stripping of effective ingredient more.
The broken technology of Chinese medicine ultra-fine powder is the new technique of prepared slices of Chinese crude drugs processing of rising along with modernized powder technology one ultrafine powder technical development, has caused the attention of manufacturer.Powder can obtain particle diameter on the one hand and reach the following powder body of 10 μ m after " micronization ", and granularity is minimum, specific surface area is multiplied, the active raising, and absorption property, surface charge, surface adhesion force all change, promote the stripping of medicine, improved utilization rate of active components; On the other hand, under this fineness condition, the cell wall breaking rate of general botanical can reach more than 95%, and intracellular effective ingredient is complete exposed state, can directly be absorbed, and reaches therapeutic effect fast and efficiently.Two aspect combineds effect, thus significantly improved drug bioavailability.
Micronizing equipment at present commonly used mainly contains three types of air-flowing type, dither formula and mechanical types, and wherein the air-flowing type super micron mill has following characteristics:
(1) degradation rate height, almost lossless;
(2) moment finishes at low temperatures in the pulverizing of powder body.Thereby the phenomenon of having avoided certain material to produce heat in crushing process and having destroyed its performance takes place;
(3) mill Jie adopts special material to make, and farthest alleviates parts wear, and almost residual the and insoluble foreign body of heavy metal free guarantees drug safety;
(4) complete requirement according to pharmacy GMP, totally-enclosed negative pressure operation, any leakage, non-environmental-pollution do not take place in powder body in crushing process.Noise is also little.
According to the literature, find through zoopery: the ultra micro of high dose seven is white to loose and can obviously resist the effect that tyrosine value, malonaldehyde value that Progesterone causes guinea pig skin, liver raise and the superoxide dismutase of skin obviously reduces, and its effect is better than with common seven of dosage and looses in vain.Find that by zoopery the yellow leaves of pulse plants the loose high, medium and low dosage of micropowder and the middle and high dosage of coarse powder all can significantly improve the carbon clearance exponential sum serum hemolysin content of normal mouse, and the mass action of micropowder obviously are better than common coarse powder.
Because superfine communication technique adopts methods such as supersonic airstream pulverize at low temperature, and is different with pure mechanical crushing method in the past, do not produce hot-spot in crushing process, and carries out under low temperature state, pulverizing speed is fast.It is diffusing to invigorate blood circulation for the exhibition muscle, and the lipid medical material accounts for more than 60% of recipe quantity, and it is big to pulverize difficulty, and very easily absorption, the moisture absorption are melted by thermal fusible, adopt superfine comminution at low temperature to cell wall breaking rate to reach more than 95%, and micronizing body tool is certain show active can, can solve problems such as above absorption moisture absorption fusion.
Compare traditional dry method micronizing, in the super micron mill of sealing, add the liquid dispersion medium of proper proportion and treat comminuting matter, the air-flow of high frequency and high-intensity positive pressure and tangential shearing force are in comminuting matter, overcome the powerful surface energy of micropowder, effectively eliminated micropowder surface air film, micropowder and medium are highly homogenized forms stable liquid system.
It is to have blood circulation promoting and blood stasis dispelling that the exhibition muscle is invigorated blood circulation diffusing, collateral dredging exhibition muscle, traditional powder of reducing swelling and alleviating pain, be used for that articular muscle due to the traumatic injury swells and ache, acute soft tissue and other chronic tissue injuries, lumbar muscle strain, joint contusion, scapulohumeral periarthritis, cervical spondylosis, lumbar disc are outstanding etc., pass through clinical application for many years and discover that the exhibition muscle is invigorated blood circulation to loose and had notable therapeutic effect for soft tissue injury pain.And through acute toxicity and long term toxicity test, show that the exhibition muscle invigorates blood circulation to loose and belong to nontoxic level medicine, do not see around the successive administration that under human dosage 100,50 and 10 multiple doses toxicity and untoward reaction take place.
The specific embodiment
According to technical scheme of the present invention, the micronizing legal system is opened up the muscle diffusing technology of invigorating blood circulation fully, adopts the preparation of low temperature wet method micronizing method, specifically comprises the following steps:
1) prescription: Radix Ginseng 125g, succinum 125g, Myrrha (processed) 125g, Olibanum (system) 125g, Sanguis Draxonis 500g, Margarita powder 125g, Radix Angelicae Sinensis 125g, Radix Notoginseng 125g, Moschus 75g, Calculus Bovis 25g;
2) will except that Moschus, Calculus Bovis, clean, wash, soak into, dry by all the other eight flavor medical materials in the prescription;
3) then at 300,000 grades of clean areas with step 2) powder medical material after the oven dry is broken, crosses sieve No. 2, obtains coarse powder;
4) with behind Moschus, Calculus Bovis and the coarse powder mixing, add 2212g PEG400, in super micron mill, pulverize 40min for 30 ℃, mean diameter is at 6.5 μ m~8 μ m; Through packing, outer package promptly.
Sanguis Draxonis is a monarch drug in the prescription, proportion is big, and removing blood stasis and relieving pain, hemostasia and promoting granulation effect are definite, it is carried out detection by quantitative can effectively control the quality of the pharmaceutical preparations, guarantee clinical efficacy, so is index with the dracorhodin, the content and the Transdermal absorption of dracorhodin that adopted high performance liquid chromatography and diffusion cell comparative study.
One, the investigation of micronizing technology
The micronizing method is carried out single factor investigate, and definite optimal processing parameter.
1, the micronizing of material is investigated
Add different liquid dispersion mediums, to the influence of micronizing effect.Get 0.5 times of recipe quantity medicinal material coarse powder, totally 4 parts, add entry, ethanol, glycerol and PEG400 respectively in the ratio of coarse powder and medium l:2, contrast with not adding the medium coarse powder, in JY92-II type super micron mill, pulverize the 30min discharging.The micropowder particle size determination: get the about 3g~5g of gained micropowder, adopt Winner99 displaing microparticle image analyzer, the granulometry scope is selected 0.6~120 μ m shelves, and parallel assay 2 times is got its meansigma methods, promptly.The results are shown in Table 1.
Table 1 variety classes disperse medium is investigated
Annotate: D
50The cumulative particle sizes percentile that is meant sample reaches 50% o'clock pairing particle diameter, also is medium particle diameter; D
90The cumulative particle sizes percentile that is meant sample reaches 90% o'clock pairing particle diameter, and the expression particle diameter accounts for 90% less than its granule.
The result shows, add disperse medium PEG400 after, material stability increases, and because of it has certain surface activity effect, forms stable hydration shell at medical surfaces, thereby increases the Transdermal absorption of fat-soluble medicine.
In the wet method micronizing, the addition of liquid dispersion medium is bigger to pulverizing result's influence, do not have effective peptizaiton very little, and material fluidity is poor, is not easy to the subsequent technique operation; Then powder uses quantitative change big too much, and therefore the additional proportion to disperse medium (90%PEG 400) has carried out preferably.The results are shown in Table 2.
Table 2 disperse medium consumption is investigated (n=2)
Result of the test shows that when medium added 1.5 ~ 2 times, material fluidity was good, and granularity is fit to.Consider from saving cost and angle easy to use, select 1.5 times of media for use.
The material thickness degree that enters super micron mill must be moderate, and granularity is too big, then is difficult for entering between mill Jie, influences crush efficiency and crushing effect; Granularity is too little, has then lost the meaning of micronizing.
Prescription: Radix Ginseng 125g, succinum 125g, Myrrha (processed) 125g, Olibanum (system) 125g, Sanguis Draxonis 500g, Margarita powder 125 g, Radix Angelicae Sinensis 125g, Radix Notoginseng 125g, Moschus 75g, Calculus Bovis 25g.
Your subclass medical material Moschus in the prescription, Calculus Bovis have been, need not coarse powder; Eight flavors such as all the other Radix Ginsengs carry out coarse powder.Get 1 times of recipe quantity medical material according to the prescription ratio, totally 4 parts, respectively with do not pulverize, standard l number sieve (10 order) crossed in coarse pulverization, No. 2, standard sieve (24 order) is crossed in coarse pulverization and coarse pulverization is crossed standard and sieved (50 order) for No. 3 and investigate.
Respectively get 500g then, place JY92-II type super micron mill to pulverize 30 minutes, investigate the micronizing effect, the results are shown in Table 3.
Table 3 material coarse pulverization degree is to the influence (n=2) of micronizing effect
Result of the test shows that along with material coarse pulverization granularity diminishes, the effect of micronizing is improved gradually.Be more or less the same with crossing 50 order coarse powder micronizing effects but cross 24 order coarse powder, so carry out micronizing again after selecting eight flavor coarse pulverizations to cross No. 2 sieves of standard.
2, the investigation of micronizing time
With the raw meal particle size is index, and the micronizing time of material is investigated, and to determine the best pulverizing time, obtains optimum crushing effect.
Test method: according to the prescription ratio, get 0.5 times of recipe quantity medical material, totally 6 parts, except that Moschus, Calculus Bovis, eight flavors such as all the other Radix Ginsengs are ground into coarse powder, cross sieve No. 2.Get eight flavor medicine coarse powder such as Moschus, Calculus Bovis and Radix Ginseng and mix, add 0.75 times of 90%PEG400, respectively at pulverizing 10min, 20min, 30min, 40in, 50min, 60min in the JY92-II type super micron mill, granularity is measured in discharging, and method is the same.The results are shown in Table 4.
The table 4 micronizing time is investigated (n=2)
Result of the test shows that along with the prolongation of micronizing time, raw meal particle size diminishes gradually.Pulverize about 40min, along with the continuation prolongation of the time of pulverizing, crushing effect almost no longer changes.So the micronizing time in this technology is decided to be 40min.
Remove 1,5,10 times medical material respectively according to the prescription ratio, will be except that Moschus, Calculus Bovis all the other eight flavors, through the medical material pre-treatment, clean, wash, soak into, dry, be ground into coarse powder earlier, cross No. 2 and sieve.The coarse powder of handling well merges Moschus, Calculus Bovis, with above ten flavor medicine mixings, adds 1.5 times, 7.5 times, 15 times PEG400 and pulverize 40min under 30 ℃ of conditions in super micron mill, investigates yield, result such as table 5:
The investigation of table 5 micronizing technology yield
Therefore, determine that micronizing exhibition muscle invigorates blood circulation diffusing preparation technology as shown in Figure 1: will be except that Moschus, Calculus Bovis all the other eight flavors, through the medical material pre-treatment, clean, with Radix Ginseng, Radix Angelicae Sinensis, Radix Notoginseng washing, soak into, oven dry, be ground into coarse powder earlier, cross No. 2 and sieve.The coarse powder of handling well merges Moschus, Calculus Bovis, with above ten flavor medicine mixings, adds PEG400, pulverizes 40min under 30 ℃ of conditions in super micron mill, and mean diameter is at 6.5 μ m~8 μ m, through packing, outer package promptly.
Annotate: pulverize coarse powder, sieve, 30 ℃ of micronizing, packing operations finish in 300,000 grades of clean areas, other operation is finished in non-clean area.
Two, the exhibition muscle Transdermal absorption research of loosing of invigorating blood circulation
1.1 the assay of dracorhodin
Measure according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 D).
Chromatographic condition and system suitability testWith the octadecylsilane chemically bonded silica is filler; With acetonitrile-0.05mol/L sodium dihydrogen phosphate (50:50) is mobile phase; The detection wavelength is 440nm; 40 ℃ of column temperatures.Number of theoretical plate calculates by the dracorhodin peak should be not less than 4000.
The preparation of reference substance solutionGet Dracoalban's reference substance 9mg, the accurate title, decide, and puts in the brown measuring bottle of 50ml, adds 3% phosphoric acid methanol solution and make dissolving, and be diluted to scale, and shaking up, precision is measured 1ml, puts in the brown measuring bottle of 5ml, add methanol to scale, shake up, promptly get (containing dracorhodin 26 μ g among every ml).
The preparation of need testing solutionThe accurate title, decided finished product 1.0g, puts in the tool plug conical flask, accurate 3% phosphoric acid methanol solution 10m1, the close plug of adding, supersound process (power 250W, frequency 30KHz) 3 minutes filters, and precision is measured subsequent filtrate 1ml, put in the brown measuring bottle of 5ml, add methanol, shake up, promptly to scale.
AlgoscopyAccurate respectively reference substance solution and each 10 μ L of need testing solution of drawing inject chromatograph of liquid, measure, promptly.
1.2 test sample preparation to be measured
No. 1: get the used exhibition muscle of the assay 1.0g that looses that invigorates blood circulation, add the 10ml propylene glycol and ground well 15 minutes, promptly.
No. 2: the exhibition muscle of getting the used micronizing of the assay 1.0g that looses that invigorates blood circulation adds the 10ml propylene glycol and ground well 15 minutes, promptly.
1.3 medicine transdermal test in vitro rate determination
1.3.1 experiment condition is determined
(1) rat skin in vitro preparation: the healthy white rat male and female have both, the about 200 ~ 250g of body weight, sacrificed by decapitation.After treatment, it is freezing skin to be put refrigerator.
(2) disperser is determined: self-control diffusion cell, effectively diffusion area 2.0cm
2, the reception tank volume is 2.5 cm
3
(3) acceptable solution: normal saline.
(4) supply with liquid: different medicinal liquid 1.5ml to be measured.
(5) experimental technique: after freezing skin thawed, be laid on the ground-in reception tank, skin inner layer is towards receiving liquid, will discharge the pond and reception tank is fixed with line.Reception tank is full of normal saline; Discharge in the pond and put different liquid to be measured.This diffusion cell is placed in the surface plate that fills water, again surface plate is placed on the constant temperature blender with magnetic force, control bath temperature (36.5 ± 0.5)
C is in the different time: 2,4,6,8,10,12 hours respectively sucking-off all receive liquid, mend full blank normal saline simultaneously and continue operation, receive all or part of evaporate to dryness of liquid, with dissolve with methanol and be moved in the tool plug test tube, measure dracorhodin content according to above-mentioned content assaying method.
Annotate: P<0.05
Find out that by experimental result and Fig. 2 after the micronizing, main limited component blood exhausts cellulose content and increases, and Transdermal absorption increases.The invigorate blood circulation Transdermal absorption effect of loosing of the exhibition muscle that micronizing is described is better than the exhibition muscle and invigorates blood circulation diffusing.
Three, the exhibition muscle diffusing pharmacodynamic study of invigorating blood circulation
Pilot project: analgesic effect test
Be subjected to the reagent thing: former exhibition muscle is invigorated blood circulation loose and the invigorate blood circulation medicated powder that looses of the exhibition muscle of micronizing adds normal saline and makes 0.2% suspension and use for subcutaneous injection respectively.
Animal subject: white mice (ICR kind system) body weight 18-22 gram, male and female half and half.
Test method and result: test tail-flick method.Get white mice and will 1/3 be dipped in 50 under the afterbody respectively
C constant temperature.In the water-bath, mensuration is because of the whipping time due to the heat, select the whipping time the white mice of 3-10 in second, every Mus is measured twice, get its meansigma methods as administration before control value, 60 qualified animals are divided into four groups at random, 15 every group, first group of feedwater 10ml/kg, give the former exhibition muscle liquid 25ml/kg that looses that invigorates blood circulation for second group, the 3rd group of exhibition muscle of giving micronizing liquid 10ml/kg that looses that invigorates blood circulation, the 4th group of exhibition muscle of the giving micronizing liquid 25ml/kg that looses that invigorates blood circulation gives indometacin 25mg/kg for the 5th group, respectively at after the administration 30,60,90, measure the whipping time in the time of 120 and 180 minutes again, deduct the whipping time before the administration, statistics is respectively organized whipping time expand, between organizing relatively (t check).Invigorate blood circulation invigorate blood circulation diffusing liquid group and the indometacin group of liquid group and the exhibition muscle of micronizing of loosing of the former exhibition muscle of result all obviously prolongs white mice because of the whipping time (seeing Table 3) due to the thermostimulation, and the exhibition muscle of micronizing invigorate blood circulation loose liquid group time expand, is maximum, and the exhibition muscle that the micronizing be described diffusing analgesic effect of invigorating blood circulation is best.
The table 3 exhibition muscle diffusing analgesic effect of invigorating blood circulation
Annotate: P<0.05
Four, " the exhibition muscle is invigorated blood circulation diffusing " treatment soft tissue injury 40 routine clinical brief summaries
Soft tissue injury is the common in daily life damages of people, belongs to " pain of injury " category of motherland's medical science.Muscle and tendon injury, the tissue injury of general reference except that fracture, dislocation, internal organs internal injury and wound, comprise tissue injurys such as skin, subcutaneous tissue, muscle, tendon, fascia, ligament, joint, nerve, blood vessel, cartilaginous opereulum, be commonly encountered diseases, the frequently-occurring disease of orthopedics department outpatient service, its because of outer by get injured by a fall, sprain and contusion, collide, sprain, extruding, the club weapons are hit and the external force effect of chronic strain etc., surmounted due to the physiological adaption of tissue, the varying in size of external force effect, injured position is different.Its pathological change, light the wounded's local organization contusion, stagnation of blood stasis, function limitation.Applicant utilization " the exhibition muscle is invigorated blood circulation diffusing " reach " the exhibition muscle of micronizing is invigorated blood circulation diffusing ", from year October in June, 2009-2009, carries out clinical observation treatment patient 40 examples, satisfactory effect, and be better than former exhibition muscle and invigorate blood circulation diffusing through the exhibition muscle of the micronizing diffusing curative effect of invigorating blood circulation.Existing brief summary is as follows:
Clinical data:This is organized among the 40 routine patients, male 16 people, women 24 people; The oldest 67 years old, minimum 5 years old, account for 32 people in 11-50 year.
The cause of disease:Get injured by a fall 8 people, break and hinder 4 people, sprain 10 people, 5 people are hindered in pinching, 4 people that strain, other 9 people.
The clinical symptoms classification:
After diagnosing, soft tissue injury person 37 people, tennis elbow 1 people, tenosynovitis 2 people.Light disease 20 people, moderate disease person 16 people, serious symptom 4 people.
Chinese medical discrimination typing: qi stagnation and blood stasis type 33 people, blood deficiency cold-condensing type 7 people.
Therapeutic Method: by the clinical symptoms classification will be light, in, severe patient is equally divided into two groups, every group 20 people.Give respectively to open up muscle invigorate blood circulation loose or the exhibition muscle of micronizing invigorate blood circulation diffusing, at injury epidermis or acupuncture point by light to the rubbing, rub of weight, 3-5 time repeatedly, every day 3 times.7 is a course of treatment, observes 2 courses of treatment altogether.
Therapeutic outcome such as following table:
Annotate: P<0.05
Brief summary:By above clinical data statistics as can be seen, by the exhibition muscle of micronizing invigorate blood circulation loose exhibition muscle that curative effect is better than former prepared invigorate blood circulation diffusing, and the exhibition muscle of two kinds of prepared invigorate blood circulation loose all easy to use, satisfactory effect.